DCL2 | Dicer-like protein 2
AS15 3100 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Arabidopsis thaliana
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Product Information
Immunogen
KLH-conjugated peptide derived from Arabidopsis thaliana DCL2 sequence, Uniprot: Q3EBC8, TAIR: AT3G03300
Host
Rabbit
Clonality
Polyclonal
Purity
Immunogen affinity purified serum in PBS pH 7.4.
Format
Lyophilized
Quantity
50 µg
Reconstitution
For reconstitution add 200 µl of sterile water
Storage
Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications
Western blot (WB)
Recommended dilution
1 : 5000-1 : 10 000 (WB)
Expected | apparent MW
156.9 | 157 kDa
Reactivity
Confirmed reactivity
Arabidopsis thaliana
Not reactive in
Nicotiana tabacum, Zea mays
Application examples
Application examples
application example
50 µg of total protein from Arabidopsis thaliana whole vegetative rosette, DCL2 overexpression line (a), wild type Col-0 (b), dcl2-1 intron insertion (c), extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with laemmli buffer at 95°C/5 min. were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent and expose to Amersham Hyperfilms ECL for 120 seconds.
Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
50 µg of total protein from Arabidopsis thaliana whole vegetative rosette, DCL2 overexpression line (a), wild type Col-0 (b), dcl2-1 intron insertion (c), extracted with extraction buffer (50 mM Tris pH7.5; 150 mM NaCl; 1 mM EDTA; 10 % v/v Glycerin; 1 mM DTT, 1x Complete Protease Inhibitor Cocktail, Roche) and denatured with laemmli buffer at 95°C/5 min. were separated on 7.5 % SDS-PAGE and blotted 1.5 h to PVDF using tank transfer. Blots were blocked with blocking buffer (5% milk powder; 1x TBS; 0.1% Tween-20) overnight at 4°C with agitation. Blot was incubated in the primary antibody at a dilution of 1:1000 for 2h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly and then washed tree times for 15 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera AS09 602) diluted to 1:20 000 in blocking buffer for 1h at RT with agitation. The blot was washed as above and developed for 5 min with chemiluminescent detection reagent and expose to Amersham Hyperfilms ECL for 120 seconds.
Courtesy of Dr. Dr. Pablo Manavella, Instituto de Agrobiotecnología del Litoral (IAL), Argentina
Additional information
Background
Background
Endoribonuclease Dicer homolog 2 is a protein which processes secondary siNAs, which is an important part in the transitive silencing of transgenes.
Alternative names: Dicer-like protein 2
Product citations
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